Effect of uncontrolled factors in a validated liquid chromatography-tandem mass spectrometry method question its use as a reference method for marine toxins: major causes for concern

Chromatographic techniques coupled to mass spectrometry is the method of choice to replace the mouse bioassay (MBA) to detect marine toxins. This paper evaluates the influence of different parameters such as toxin solvents, mass spectrometric detection method, mobile-phase-solvent brands and equipment on okadaic acid (OA), dinophysistoxin-1 (DTX-1), and dinophysistoxin-2 (DTX-2) quantification. In addition, the study compares the results obtained when a toxin is quantified against its own calibration curve and with the calibration curve of the other analogues. The experiments were performed by liquid chromatography (LC) and ultraperformance liquid chromatography (UPLC) with tandem mass spectrometry detection (MS/MS). Three acetonitrile brands and two toxin solvents were employed, and three mass spectrometry detection methods were checked. One method that contains the transitions for azaspiracid-1 (AZA-1), azaspiracid-2 (AZA-2), azaspiracid-3(AZA-3), gimnodimine (GYM), 13-desmethyl spirolide C (SPX-1), pectenotoxin-2 (PTX-2), OA, DTX-1, DTX-2, yessotoxin (YTX), homoYTX, and 45-OH-YTX was compared in both instruments. This method operated in simultaneous positive and negative ionization mode. The other two mass methods operated only in negative ionization mode, one contains transitions to detect DTX-1, OA DTX-2, YTX, homoYTX, and 45-OH-YTX and the other only the transitions for the toxins under study OA, DTX-1, and DTX-2. With dependence on the equipment and mobile phase used, the amount of toxin quantified can be overestimated or underestimated, up to 44% for OA, 46% for DTX-1, and 48% for DTX-2. In addition, when a toxin was quantified using the calibration curve of the other analogues, the toxin amount obtained is different. The maximum variability was obtained when DTX-2 was quantified using either OA or a DTX-1 calibration curve. In this case, the overestimation was up to 88% using the OA calibration curve and up to 204% using the DTX-1 calibration curve. In summary, the correct quantification of DSP toxins by MS detection depends on multiple factors. Since these factors are not taken into account in a validated protocol, these results question the convenience of having MS/MS as a reference method for protecting consumers of marine toxins, moreover if toxicity of each group is considered independently and total toxicity is not summed anymore as it is in the MBA.     

杰昆·索罗拉火车站,巴伦西亚,西班牙

何谓永恒城市与火车之间的爱恨情仇/关系源远流长,有过激情四溢的乐章,也有过痛苦折磨的片断,有过甜蜜美好的瞬间,也有公诸于众的背叛。而今,两者之间的关系历经变化,今非昔比。然而在巴伦西亚的杰昆·索罗拉火车站,城市与火车之间的激情烈焰仍在燃烧。新车站在允许高速火车进行停靠的同时,完成了将铁轨迁入地下的工程:     

Curcumin Sensitises Cancerous Kidney Cells to TRAIL Induced Apoptosis via Let-7C Mediated Deregulation of Cell Cycle Proteins and Cellular Metabolism

Targeted therapies are the most attractive options in the treatment of different tumours, including kidney cancers. Such therapies have entered a golden era due to advancements in research, breakthroughs in scientific knowledge, and a better understanding of cancer therapy mechanisms, which significantly improve the survival rates and life expectancy of patients. The use of tumour necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL) as an anticancer therapy has attracted the attention of the scientific community and created great excitement due to its selectivity in targeting cancerous cells with no toxic impacts on normal tissues. However, clinical studies disappointingly showed the emergence of resistance against TRAIL. This study aimed to employ curcumin to sensitise TRAIL-resistant kidney cancerous ACHN cells, as well as to gain insight into the molecular mechanisms of TRAIL sensitization. Curcumin deregulated the expression of apoptosis-regulating micro Ribonucleic Acid (miRNAs), most notably, let-7C. Transfecting ACHN cells with a let-7C antagomir significantly increased the expression of several cell cycle protein, namely beta (β)-catenin, cyclin dependent kinase (CDK)1/2/4/6 and cyclin B/D. Further, it overexpressed the expression of the two key glycolysis regulating proteins including hypoxia-inducible factor 1-alpha (HIF-1α) and pyruvate dehydrogenase kinase 1 (PDK1). Curcumin also suppressed the expression of the overexpressed proteins when added to the antagomir transfected cells. Overall, curcumin targeted ACHN cell cycle and cellular metabolism by promoting the differential expression of let-7C. To the best of our knowledge, this is the first study to mechanistically report the cancer chemosensitisation potential of curcumin in kidney cancer cells via induction of let-7C.     

11C-Methionine PET/CT in Assessment of Multiple Myeloma Patients: Comparison to 18F-FDG PET/CT and Prognostic Value

Multiple myeloma (MM) is the second most common haematological malignancy and remains incurable despite therapeutic advances. ¹⁸F-FDG (FDG) PET/CT is a relevant tool MM for staging and it is the reference imaging technique for treatment evaluation. However, it has limitations, and investigation of other PET tracers is required. Preliminary results with L-methyl-[¹¹C]- methionine (MET), suggest higher sensitivity than ¹⁸F-FDG. This study aimed to compare the diagnostic accuracy and prognostic value of ¹FDG and MET in MM patients. We prospectively compared FDG and MET PET/CT for assessment of bone disease and extramedullary disease (EMD) in a series of 52 consecutive patients (8 smoldering MM, 18 newly diagnosed MM and 26 relapsed MM patients). Bone marrow (BM) uptake patterns and the detection of focal lesions (FLs) and EMD were compared. Furthermore, FDG PET parameters with known MM prognostic value were explored for both tracers, as well as total lesion MET uptake (TLMU). Median patient age was 61 years (range, 37–83 years), 54% were male, 13% of them were in stage ISS (International Staging System) III, and 31% had high-risk cytogenetics. FDG PET/CT did not detect active disease in 6 patients, while they were shown to be positive by MET PET/CT. Additionally, MET PET/CT identified a higher number of FLs than FDG in more than half of the patients (63%). For prognostication we focussed on the relapsed cohort, due to the low number of progressions in the two other cohorts. Upon using FDG PET/CT in relapsed patients, the presence of more than 3 FLs (HR 4.61, p = 0.056), more than 10 FLs (HR 5.65, p = 0.013), total metabolic tumor volume (TMTV) p50 (HR 4.91, p = 0.049) or TMTV p75 (HR 5.32, p = 0.016) were associated with adverse prognosis. In MET PET/CT analysis, TMTV p50 (HR 4.71, p = 0.056), TMTV p75 (HR 6.27, p = 0.007), TLMU p50 (HR 8.8, p = 0.04) and TLMU p75 (HR 6.3, p = 0.007) adversely affected PFS. This study confirmed the diagnostic and prognostic value of FDG in MM. In addition, it highlights that MET has higher sensitivity than FDG PET/CT for detection of myeloma lesions, including FLs. Moreover, we show, for the first time, the prognostic value of TMTV and TLMU MET PET/CT in the imaging evaluation of MM patients.     

A Compact Four-Quadrant Floating-Gate MOS Multiplier

A compact, four-quadrant analog CMOS multiplier featuring wide dynamic range is presented. The capacitive voltage division obtained by the use of Floating-Gate MOS (FGMOS) transistors, and an accurate wide-swing current mirror based on active bootstrapping, allow a wide input range, low harmonic distortion, and high linearity. Simulation and measurement results for a 0.8 μm CMOS prototype demonstrate the validity of the proposed approach.     

A macrokinetic approach to crystallization applied to a new thermoplastic polyimide (New TPI) as a model polymer

The application of a new macrokinetic approach to polymer crystallization is applied in this work to a new thermoplastic polyimide (New TPI) recently developed as a matrix of advanced polymer matrix composites. The slow crystallization kinetics presented by New TPI in the entire range between the glass-transition temperature and the melting point makes TPI an excellent model polymer for testing crystallization models. In the approach presented here the variation of the induction time with the temperature is included in the Nakamura model for nonisothermal crystallization, and a simplified expression of the kinetic constant as a function of the temperature is adopted. The proposed model is verified through a comparison with a complete set of experimental data, ranging from the melting point to the glass-transition temperature of the polymer, obtained in isothermal and nonisothermal conditions. Moreover phase transformation diagrams (TTT and CCT plots) are presented, providing a fundamental tool for understanding the crystallization behavior of semicrystalline matrices and to determine the more appropriate processing conditions. © 1995 John Wiley & Sons, Inc.     

Role of Bioflocculation on Chemical Oxygen Demand Removal in Solids Contact Chamber of Trickling Filter/Solids Contact Process

The trickling filter/solids contact (TF/SC) process was developed in the late 1970s to improve the quality of the final effluent from existing trickling filter plants, to be able to meet stricter Environmental Protection Agency effluent requirements. Although this process has successfully achieved this objective, it is still not completely understood, there is limited information regarding the flocculation phenomena occurring in the solids contact chamber (SCC), and no information could be found on the relationship between flocculation and organic matter removal kinetics. To better understand the kinetics of biological flocculation in a continuous flow SCC, a long-term experimental program was conducted using a TF/SC pilot plant constructed at the Marrero, La., wastewater treatment plant. This program started in January 1998 and has continued through date. The present article will focus on two major areas: (1) the kinetics of bioflocculation in the SCC; and (2) effect of bioflocculation on chemical oxygen demand (COD) removal. Analysis of the wastewater composition revealed that, on the average, only 18.7% of the total COD in the SCC influent is truly dissolved. Therefore, most of the total COD removal observed in the SCC must be due to a physical process, such as flocculation. The experimental data confirmed that flocculation of the particulate COD contained in the trickling filter effluent explains the high total COD removal observed at the SCC. Both total and colloidal COD removals are well explained by the first-order flocculation model.     

Very Low Voltage MOS Translinear Loops Based on Flipped Voltage Followers

A novel technique for operating MOS Translinear loops at very low supply voltages is described, based on the use of Flipped Voltage Followers for biasing the loops. The resulting topologies, suited to standard CMOS processes, can be successfully applied to a varied repertory of low-voltage analog circuits, such as squarers, multipliers, filters, oscillators, and RMS-DC converters. Measurement results for a geometric-mean and a squarer/divider circuit demonstrate on silicon the usefulness of this technique.     

Flavonoids as stabilizers of fish oil: An alternative to synthetic antioxidants

The antioxidant activities against fish oil oxidation of six commercially available flavonoids and of five flavonoids purified from two Chilean native plants were compared to those ofdl-α-tocopherol and of two synthetic antioxidants, butylated hydroxytoluene and butylated hydroxyanisole. Among the commercial flavonoids, catechin, morin and quercetin showed a higher activity when fish oil oxidation (either spontaneous or Fe²⁺-induced) was assessed from the formation of peroxides or thiobarbituric acid-reactive substances. Among the native flavonoids, the 5,3′,4′-trihydroxy-7-methoxy flavanone (designated as Pt-2) showed the highest antioxidant activity. Mixtures of quercetin or of Pt-2 withdl-α-tocopherol produced better inhibitory effects when compared to that of each substance assayed by itself. Also, when Pt-2 and quercetin were assayed in combination (0.3 g/kg oil and 0.7 g/kg oil, respectively), a synergistic antioxidant effect was observed. Results indicate that several flavonoids could be used as natural antioxidants as a means to replace those synthetic antioxidants, the use of which has been questioned.     

Validation of the rancimat test for the assessment of the relative stability of fish oils

The induction periods for the peroxidation of various fish oils at 55–90°C were studied by the Rancimat test. The natural logarithms of the induction periods varied linearly with respect to temperature, with a mean coefficient of −7.5×10⁻²°C⁻¹, which was significantly different from that reported for vegetable oils. The activation energy for the formation of volatile acids had a mean value of 38.9 kJ/mol and was independent of the fish oil source. Peroxide formation under Rancimat test conditions followed first-order kinetics. The same kinetics were followed under Schaal Oven test conditions (forced-air oven, 60°C). On the basis of the results obtained, the Rancimat test appears to be useful in determining the relative stabilities of fish oils without the change in peroxide decomposition kinetics that may occur at elevated temperatures.